Silkworm larvae plasma (SLP) assay for detection of bacteria: False positives secondary to inflammation in vivo

Ma, Michelle; Rice, Tyler; Percopo, Caroline M.; Rosenberg, Helene F.

doi:10.1016/j.mimet.2016.11.009

Cited by 4 publications

(6 citation statements)

References 26 publications

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“…administration. Ma et al [19] also reported rapid clearance of gDNA from both live and inactivated Lp.…”

Section: Results and Dicussionmentioning

confidence: 94%

“…Detection of Lp genomic (g)DNA in lung tissue was performed as described [19]. Lungs from mice inoculated with Lp were stored at 280°C in RNAlater (Thermo Fisher Scientific); gDNA was isolated and evaluated on an ABI 7500 Real Time PCR system (Thermo Fisher Scientific), with primers and probe designed to detect the Lp 16s-23s intergenic region as described [20], including forward primer, 59-TGG ATC ACC TCC TTT CTA AGG AAT-39; reverse primer, 59-TGT TCT CGG TTT CAT TAT GAA AAA ATA-39; and probe, 59-ACA TTC TTC GAA ACT TTG T-39-MGB and calibrated via interpolation to standard curves.…”

Section: Detection Of Lp Genomic Dnamentioning

confidence: 99%

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Administration of immunobiotic Lactobacillus plantarum delays but does not prevent lethal pneumovirus infection in Rag1−/− mice

Percopo

Rosenberg

2017

Journal of Leukocyte Biology

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Administration of immunobiotic (Lp) directly to the respiratory mucosa promotes cross-protection against lethal pneumovirus infection via B-cell-independent mechanisms. In this study, we examined Lp-mediated cross protection in mice which cannot clear virus from lung tissue. Although Lp was initially protective, mice ultimately succumbed to a delayed lethal outcome associated with local production of the proinflammatory cytokines CCL1, -2, and -7, granulocyte recruitment, and ongoing virus replication. By contrast, CD8 mice, which are fully capable of clearing virus, are protected by Lp with no delayed lethal outcome, granulocyte recruitment to the airways, or induction of CCL7. Repeated administration of Lp to virus-infected mice had no impact on delayed mortality. Moreover, administration of Lp to the respiratory mucosa resulted in no induction of IFN-α or -β in or wild-type mice, and IFNR gene deletion had no impact on Lp-mediated protection. Overall, our findings indicate that although Lp administered to the respiratory tract has substantial impact on lethal virus-induced inflammation in situ, endogenous virus clearance mechanisms are needed to promote sustained protection. Our results suggest that a larger understanding of the mechanisms and mediators that limit acute virus-induced inflammation may yield new and useful therapeutic modalities.

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“…administration. Ma et al [19] also reported rapid clearance of gDNA from both live and inactivated Lp.…”

Section: Results and Dicussionmentioning

confidence: 94%

Section: Detection Of Lp Genomic Dnamentioning

confidence: 99%

Administration of immunobiotic Lactobacillus plantarum delays but does not prevent lethal pneumovirus infection in Rag1−/− mice

Percopo

Rosenberg

2017

Journal of Leukocyte Biology

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“…For example, in the above-mentioned study by Zhiping et al [14], the exposure to peptidoglycans was assessed using a muramic acid as its marker. Also, recent reports suggest that the SLP test should be used with caution as it can give the "false-positive results" contributing to overestimation of the real exposure [30].…”

Section: Discussionmentioning

confidence: 99%

Across-shift changes in upper airways after exposure to bacterial cell wall components

Cyprowski¹,

Stobnicka-Kupiec²,

Górny³

et al. 2019

Ann Agric Environ Med.

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Introduction. To assess the across-shift changes of cytokine concentrations in nasal lavage (NAL) samples were collected from workers exposed to bacterial cell wall components present in organic dust in three different occupational environments. Materials and method. The study was conducted in 38 employees including 10 workers from a municipal waste sorting plant (WSP), 20 from a sewage treatment plant (STP) and 8 from an office building (OB), who were established as a reference group, not exposed to organic dust. Interleukins 1β (IL-1β), 6 (IL-6), 8 (IL-8) and tumour necrosis factor alpha (TNF-α) were examined in NAL before and after work shift on Wednesdays. Bioaerosol exposure was determined by personal measurements and analysed for organic dust, endotoxins (END) and peptidoglycans (PGN). Results. The analysis included the results for IL-8 only, because for the other cytokines their concentrations in 80% of cases were below the detection level. The most polluted were the workplaces in WSP with average concentrations of organic dust -3.47 mg/m 3 , END -96.31 ng/m 3 and PGN -571.88 ng/m 3 . The results of IL-8 showed a significant difference between the studied groups after the work shift (p=0.007). Among WSP workers concentrations of IL-8 increased also significantly (p=0.015) during the work shift. Multivariate analysis showed that organic dust and END were the factors that in the most distinct way (p<0.001) influenced changes of IL-8 levels in NAL. Conclusions. Each alteration in the composition of bioaerosols will probably determine the changes in the mechanisms responsible for both formation and modulation of inflammatory reactions in exposed workers.

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“…This discrepancy could however be due to differences in PGN reactivity. The SLP reagent was indeed reported to react with free PGN, but also with trace of living or inactivated bacteria [29] , [38] . In contrast, the size of PGN fragments was shown to determine the responses triggered by inflammatory cells in human and animal models.…”

Section: Discussionmentioning

confidence: 99%

“…Unfortunately, PGN detection in icodextrin batches is difficult, because of its relatively low concentration by comparison with the icodextrin itself, which behaved as an interfering substance. Moreover, routine monitoring for PGN with SLP assay has some shortcomings, because of limitation in the detection [38] and cross-reactivity with other substances, such like β1-3 glucans [29] . In the light of needs for other tests, we decided to compare the efficacy of different bio-assays to detect bacterial contamination in glucose polymer samples.…”

Section: Discussionmentioning

confidence: 99%

Use of Toll-like receptor assays for the detection of bacterial contaminations in icodextrin batches released for peritoneal dialysis

et al. 2017

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Silkworm larvae plasma (SLP) assay for detection of bacteria: False positives secondary to inflammation in vivo

Abstract: The silkworm larvae plasma (SLP) assay has been developed as a means to detect bacterial peptidoglycan as a surrogate for live bacteria. Here, we present results that indicate that generation of melanin by this assay is not fully reliable as a surrogate marker for bacterial count.

Cited by 4 publications

References 26 publications

Administration of immunobiotic Lactobacillus plantarum delays but does not prevent lethal pneumovirus infection in Rag1−/− mice

Administration of immunobiotic Lactobacillus plantarum delays but does not prevent lethal pneumovirus infection in Rag1−/− mice

Across-shift changes in upper airways after exposure to bacterial cell wall components

Use of Toll-like receptor assays for the detection of bacterial contaminations in icodextrin batches released for peritoneal dialysis

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